Integrated miRNA-cytokine profiling reveals a molecular signature of cachexia in patients with chronic heart failure

Figures

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  Figure 1. Circulating miRNAs and cytokines as molecular signatures of heart failure. Differences in circulating miRNA levels between CHF patients and controls. Circulating miRNA levels were analyzed by quantitative Real-Time PCR from patients with heart failure (HF) (n = 25) and control group (n = 10). Data shows significant downregulation of miR-15b (P < .001) (A), −29b (P < .001) (D), −133a (P = .005) (E) and −486 (P < .001) (F) in CHF vs controls. No statistical significance was observed for miR-21 (B) and −29a (C) levels. Data were normalized to miR-16 from two biological replicates, with a calibrator used as the internal control. Data are expressed as fold change and are presented as mean ± SEM. Student’s t-test or Mann–Whitney test was used for normally distributed data or non-normally distributed data, respectively. ** P ≤ .01, *** P ≤ .001.

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  Figure 2. Circulating miRNAs and cytokines as molecular signatures of heart failure. Differences in serum levels of GDF-15 and FGF-21 in CHF patients and controls. Serum levels of GDF-15 by ELISA assay in patients with heart failure (HF) (n = 25) and the control group (n = 10). (A) Data showed significant upregulation of GDF-15 in CHF vs controls (P < .001). (B) FGF-21 levels did not differ significantly between groups. Correlations between GDF-15 and selected miRNAs in CHF patients. (C, D) In CHF patients, GDF-15 levels were positively associated with miR-29b (ρ = 0.42, P = .037) and miR-486 (ρ = 0.42, P = .039). Data are expressed as fold change and are presented as mean ± SEM. Student's t-test or Mann–Whitney test was used for normally distributed data or non-normally distributed data, respectively. *** P ≤ .001.

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  Figure 3. Molecular profiles in CHF patients. Differences in circulating miRNA levels among heart failure patients with and without cardiac cachexia. Circulating miRNA levels were analyzed in CHF patients with cardiac cachexia (CC) (n = 10) and without cardiac cachexia (nCC) (n = 15), and a control group (n = 10). (D) Data show significant downregulation of miR-29b (P = .019) in HF with CC vs nCC and vs controls (P = .001). Significant modulations were observed between CHF patients with CC or nCC vs controls for miR-15b (A), −133a (E), and −486 (F) (all P < .05). No other statistical significances were observed for miR-21 (B), and −29a (C). Data are expressed as fold change and are presented as mean ± SEM. One-way ANOVA test or Kruskal–Wallis test was used for normally distributed data or non-normally distributed data, respectively. * P < .05, ** P ≤ .01, *** P ≤ .001.

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  Figure 4. Molecular profiles in CHF patients. Differences in serum levels of GDF-15 and FGF-21 among heart failure patients with and without cardiac cachexia. Circulating levels of GDF-15 and FGF-21 were analyzed in CHF patients with cardiac cachexia (CC) (n = 10) and without cardiac cachexia (nCC) (n = 15), and a control group (n = 10). (A) Significant difference in GDF-15 levels was observed between patients with CC or nCC vs controls (all P < .001). (B) Levels of FGF-21 were significantly higher in CHF with CC compared with nCC (P = .012) and vs controls (P = .010) (C). Spearman’s correlation analysis revealed that in CHF patients with CC, GDF-15 negatively correlated with miR-21 (ρ = −0.81, P = .007). (D) In the same group, FGF-21 negatively correlated with miR-486 (ρ = −0.69, P = .035). Data are expressed as fold change and are presented as mean ± SEM. One-way ANOVA test or Kruskal–Wallis test was used for normally distributed data or non-normally distributed data, respectively. * P < .05, *** P ≤ .001.

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  Figure 5. Molecular profiles in cachectic patients with severe CHF. Circulating miRNA levels and GDF-15 correlations according to ejection fraction and cachexia status in CHF patients. Circulating miRNA levels were analyzed in CHF patients with preserved ejection fraction without cachexia (HFpEF + nCC) (n = 7) and with reduced ejection fraction with cachexia (HFrEF + CC) (n = 5) and a control group (n = 10). (A) Data shows significant downregulation of miR-29b (P = .019) in HFrEF + CC vs HFpEF + nCC and vs controls (P = .002). (B) In HFpEF + nCC, GDF-15 showed a strong positive correlation with miR-29b (r = 0.82, P = .034). Data are expressed as fold change and are presented as mean ± SEM. One-way ANOVA test or Kruskal–Wallis test was used for normally distributed data or non-normally distributed data, respectively. * P < .05, ** P ≤ .01.

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  Figure 6. Molecular profiles in cachectic patients with severe CHF. FGF-21 concentrations and their associations with NYHA functional class in patients with CHF. (A) Data show higher FGF-21 levels in NYHA class III-IV (n = 13) compared with class I-II (n = 9) (P = .050) and the control group (n = 10) (P = .050). (B) In NYHA class III-IV, FGF-21 positively correlated with miR-15b (r = 0.68, P = .014). Data are expressed as fold change and are presented as mean ± SEM. One-way ANOVA test or Kruskal–Wallis test was used for normally distributed data or non-normally distributed data, respectively. * P < .05.